Ankit was born in Kanpur, Uttar Pradesh. His parents own a music troupe in Kanpur called "Raju Suman and Party" which performs at religious convocations. His mother, is a devotional singer. Though his parents wanted him to complete his studies first, his inclination towards music led them to decide on training him professionally. He trained in classical music from Vinod Kumar Dwivedi before training in piano, Dhrupad and Western vocals. He instituted at and won several local music competitions during the time. He used to work as a production head at a radio station in Gwalior, before he moved to Mumbai in December 2007 along with his brother Ankur Tiwari.
In Mumbai, Tiwari met Pradeep Sarkar who was working on a film at that time. Though the film got shelved, Sarkar gave him, along with other musicians, a chance to work on jingles. It was then he composed a couple of background scores for television programmes. Tiwari met Habib Faisal, who offered him to compose the music for his directorial debut film Do Dooni Chaar which was released in 2010. Later for producer Tigmanshu Dhulia he composed the theme song of Saheb, Biwi Aur Gangster (2011). With this film, Tiwari made his singing debut where he rendered the song "Saheb Bada Hathila" along with Vipin Aneja.
In 2012, Tiwari narrated the song "Sunn Raha Hai" to Mahesh Bhatt and Mohit Suri, the director of the film Aashiqui 2. The song was also recorded in his voice and the female-version was rendered by Shreya Ghoshal. Apart from receiving critical acclaim, the song fetched Tiwari with multiple awards and nominations. He shared his first Filmfare Award as the Best Music Director along with Mithoon and Jeet Gannguli. The team received the Best Music Director Award from other ceremonies including IIFA Awards and Zee Cine Awards. Apart from music direction, Tiwari received his second nomination in Filmfare Awards as the Best Male Playback Singer. As his second release of the year, Tiwari collaborated with Sachin Gupta—marking his first song to the tunes of another composer—for the song "Aag Ka Dariya" from Issaq.
2014 began with the film Dee Saturday Night, where two of his songs, "Falsafa Mera Falsafa" and "Jhatak Ke Nacho" went mostly unnoticed with the music listeners. He then performed the track "Kaisa Hai Dard Mera" which is composed by Ismail Darbar for Subhash Ghai's Kaanchi. Tiwari composed two tracks for the film Samrat & Co., titled "Sawaalon Mein" and a party song "Tequila Wakila" where both are rendered by Ghoshal. During the year, he performed three tracks for two of Himesh Reshammiya's films, The Xposé and Action Jackson. His rendition for the song "Sheeshe Ka Samundar" from the former received critical acclaim, while "Punjabi Mast" and "Dhoom Dhaam" from the latter received negative response from critics. In July 2014, a duet with Tulsi Kumar titled "Kuch Toh Hua Hai" was released included in the film Singham Returns. The year ended with "Dil Darbadar" from PK. Apart from all the songs released during the year, the most successful track was "Galliyan" from Ek Villain. The song was a critical and commercial success; Mohar Basu from Koimoi described the song as "the most attractive song of the album" stating "Tiwari's music and voice adds the required finesse in this song and what he has made darn great!". The song fetched him several awards for Best Male Playback Singer including from Filmfare, Star Guild and Stardust Awards. Subsequently he received his fourth nomination in Filmfare as the Best Music Director Award at 60th Britannia Filmfare Awards. In the year, Tiwari made his debut in a Kannada film with the song "Nillu Nillu" from Dil Rangeela composed by Arjun Janya.
Tiwari composed and sung the song "Katra Katra" along with Prakriti Kakar for the film Alone (2015). They again collaborated for the song "Bheegh Loon" from Khamoshiyan. He then composed three tracks for the film Roy, "Tu Hai Ki Nahi", "Yaara Re" which is rendered by KK and "Boond Boond" performed by Tiwari himself. The unplugged version of "Tu Hai Ki Nahi" performed by Tulsi Kumar along with the original received mixed reactions from critics though infusion of the whistle's tune was appreciated.
Tiwari stated that he wants to explore different genres given the right opportunity, though it shouldn't be vulgar and should hold meaning. Satyajit from Glamsham called his second composed song "Saheb Bada Hatila" from Saheb, Biwi Aur Gangster a "rock-musical" arrangement. In his next composition "Sunn Raha Hai", the male version features electronic distortion guitar, zitar and drums while the female version is supported by instruments like ghatam, flute, santoor and acoustic guitar. The variance in composition was particularly applauded by critics.
In 2014, Tiwari composed "Tequila Wakila" which lies in Spanish style musical background settings of bass, drums, saxophone, trumpets and guitar while "Sawalon Mein" from the same film has a mysterious type semi-philosophical composition which is decorated with haunting piano notes and bell sounds in the background. Rajiv Vijayakar from Bollywood Hungama mentioned that Tiwari seems to be "evolving into a composer with a decent range". In the composition of the song "Galliyan" it transforms from "being slow-paced, to quickly accelerating into heavy rock music and then settling into soothing traditional Indian-themed sounds", while the unplugged version dominantly uses the acoustic guitar alongside piano. However for his next song "Kuch Toh Hua Hai", Surabhi Redkar for Koimoi commented; "Experimentation wise, the song shows no novelty as the musical arrangements are not peculiarly impressive". According to reviewers, his last song released in 2014, "Dil Darbadar" has a huge "Galliyan" hangover and "doesn't actually fit into the scheme of things". In September 2014, Tiwari said; "People usually call me for love songs or songs of separation, which I love composing. But I don't want to restrict myself to a particular genre".
"Katra Katra" from 2015 had a similar fate with "Dil Darbadar" as the musical arrangements of both the song are similar to his earlier ventures. Redkar noted; "If only Tiwari could come up with something new [...] the only problem is that it all sounds the same". His next was a passionate and intense composition, "Bheegh Loon" which fits with the erotic thriller mood of the film. In "Tu Hai Ki Nahin" from Roy Tiwari used minimal musical arrangements mainly focusing on vocals and incorporate a whistle—first time he has used whistle as an instrument—which according to music listeners "sounds brilliant and has high recall value", while the next two tracks "Boond Boond" and "Yaara Re" were called "experimental" in terms of arrangement.
Tiwari named the singers like KK, Puerto Rican and Ricky Martin as his idols. He picked Nusrat Fateh Ali Khan as one of his inspiration and dedicated a song to him from one of his composition at MTV Unplugged 4 – Episode 06. Among the actors, Tiwari expressed his desire to dub for Shah Rukh Khan and Ranbir Kapoor.
Popular singer Rahat Fateh Ali Khan mentioned that he likes Tiwari's singing along with Arijit Singh's.
Tiwari's early two releases "Sunn Raha Hai" and "Aag Ka Dariya" represent his "young energetic and strong vocals" which was well received by the critics. "Kaisa Hai Dard Mera", despite with the similar pattern has a semi classical touch in his rendition. His one of the most successful venture "Galliyan" provided Tiwari with a high vocal range which was particularly appraised in the song though Bollywood Hungama wrote that he sings the track in a "gimmicky tenor that sounds fake". His voice and rendition was criticised in "Dil Darbadar" where IBNLive wrote; "Tiwari's choice to distort vocals using the Auto-tune software takes away the charm and rawness of a plaintive melody like "Dil Darbadar".
On 9 May 2014, Tiwari was arrested for allegedly raping his girlfriend, while his brother was booked on complaint of making threats to her life. Tiwari was apprehended under rape charges after a 28-year-old advertising professional approached the Versova police station alleging rape. In her statement, she said that he raped her on many occasions since July 2013, after promising marriage. A week later, he was granted bail by the Sessions Court on the condition that he furbish a bond of Rs 20,000.
On April 2017, Tiwari and his brother were acquitted (freed) from all charges by the Court. As the prosecution failed to gather enough evidence against him and his brother.
After getting acquitted he spoke to the Times of India, he said:
Justice has finally been done. I knew that God and truth were on my side, and that I had done no wrong. Finally, everyone knows the truth. I can never adequately thank the court and the people who helped me through this ordeal.For three and half years my family and I suffered through this ordeal.